Abstract: This experiment chose the royal jelly from Heilongjiang Mishan as raw material and took water-soluble protein extraction rate as evaluation indicator.Based on single factor experiment and Plackett-Burman design, the response surface optimization method was used to optimize the extraction of water-soluble protein from royal jelly by water extraction ammonium sulfate precipitation.The experiment showed that the optimum technological parameters were material water mass ratio 1 ∶ 20, ultrasonic extraction time 40 minutes, ammonium sulfate saturation 89%, pH5, precipitation time 50 minutes.Under the condition of this, the water-soluble protein extraction rate of royal jelly was up to 80.22%, this method of experiment was the better way to extract water-soluble protein of royal jelly.